Overview

  • Product nameAnti-P2X3 antibody
    See all P2X3 primary antibodies
  • Description
    Rabbit polyclonal to P2X3
  • Tested applicationsSuitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Pig, Zebrafish
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 250 - 350 of Human P2X3.

    (Peptide available as ab94545.)

  • Positive control
    • This antibody gave a positive signal in human testis tissue lysate and in the following whole cell lysates: Jurkat; HepG2.

Properties

Applications

Our Abpromise guarantee covers the use of ab75453 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 49 kDa (predicted molecular weight: 44 kDa).

Target

Anti-P2X3 antibody images

  • All lanes : Anti-P2X3 antibody (ab75453) at 1 µg/ml

    Lane 1 : Human testis tissue lysate - total protein (ab30257)
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 44 kDa
    Observed band size : 49 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 12 kDa,18 kDa,78 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute
  • ICC/IF image of ab75453 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75453, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and MCF7 cells at 5µg/ml.
  • IHC image of P2X3 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab75453, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX

References for Anti-P2X3 antibody (ab75453)

ab75453 has not yet been referenced specifically in any publications.

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